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Table 2 Primer sets for PCR amplification of FVBD agents

From: Bacterial and protozoal agents of feline vector-borne diseases in domestic and stray cats from southern Portugal

Pathogen Gene Primers Product size (bp) PCR conditions Reference
Anaplasma spp./Ehrlichia spp. 16S rRNA EHR16SD: 5′-GGT ACC YAC AGA AGA AGTCC-3′ 345 95°C, 5 min; 35 cycles [94°C 30 sec, 55°C 30 sec, 72°C 90 sec]; 72°C, 5 min [12]
  EHR16SR: 5′-TAG CAC TCA TCG TTT ACAGC-3′
Babesia spp. 18S rRNA PIRO-A: 5′-AAT ACC CAA TCC TGA CACAGG G-3′ 400 95°C, 5 min; 35 cycles [94°C 30 sec, 55°C 30 sec, 72°C 90 sec]; 72°C, 5 min [13]
  PIRO-B: 5′-TTA AAT ACG AAT GCC CCCAAC-3′
Bartonella spp. 16S-23S rRNA 325 s: 5′-CTTCAGATGATGATCCCAAGCCTTCTGGCG-3′ 500-800 95°C, 5 min; 55 cycles [95°C 15 sec, 66°C 15 sec, 72°C 15 sec]; 72°C, 1 min [14]
  1100as: 5′-GAACCGACGACCCCCTGCTTGCAAAGCA-3′
B. burgdorferi s.l. 5S-23S rRNA Outer primers: 380   [15]
  23SN1: 5′-ACCATAGACTCTTATTACTTTGAC-3′ 94.5°C, 1 min; 25 cycles [94°C 30 sec, 52°C 30 sec, 72°C 1 min]; 72°C, 5 min
  23SC1: 5′-TAAGCTGACTAATACTAATTACCC-3′
  Inner primers: 225  
  23SN2: 5′-ACCATAGACTCTTATTACTTTGACCA-3′ 94.5°C, 1 min; 40 cycles [94°C 30 sec, 55°C 30 sec, 72°C 1 min]; 72°C, 5 min
  5SCB: 5′-biotin-GAGAGTAGGTTATTGCCAGGG-3′
Hepatozoon spp. 18S rRNA HEP-F: 5′-ATA CAT GAG CAA AAT CTC AAC-3′ 626-666 95 °C, 5 min; 34 cycles [95 °C 20 sec, 55 °C, 30 sec, 72 °C, 90 sec]; 72 °C, 5 min [16]
  HEP-R: 5′-CTT ATT ATT CCA TGC TGC AG-3′
Leishmania spp. Small subunit rRNA Outer primers: 603   [17]
  R221: 5′-GGTTCCTTTCCTGATTTACG-3′ 94°C, 5 min; 35 cycles [94°C 30 sec, 60°C 30 sec, 72°C 30 sec]; 72°C, 10 min
  R332: 5′-GGCCGGTAAAGGCCGAATAG-3′
  Inner primers: 358   [18]
  R223: 5′-TCCCATCGCAACCTCGGTT-3′ 94°C, 5 min; 35 cycles [94°C 30 sec, 65°C 30 sec, 72°C 30 sec]; 72°C, 10 min
   R333: 5′-AAAGCGGGCGCGGTGCTG-3′